翘嘴鳜生长激素cDNA克隆及其真核表达载体的构建

Cloning of Siniperca chuatsi growth hormone cDNA and construction of its eukaryotic expression vector

根据GenBank上大眼鳜生长激素cDNA序列(收录号:AY155227)设计了1对特异性引物,应用RT-PCR技术从翘嘴鳜(Siniperca chuatsi)脑垂体中克隆了生长激素基因(scGH)编码区序列并对其真核表达载体的构建进行了研究。结果显示:扩增片段全长615 bp,共编码204个氨基酸。该序列与GenBank上大眼鳜生长激素cDNA序列同源性为99.84%。将scGHcDNA序列定向插入pYES2/CT真核表达载体中,经过筛选、酶切和测序,证明重组子中确实插入了翘嘴鳜GH片段。

One special primer was designed based on the Siniperca chuatsi growth hormone cDNA order in the GenBank （embody number： AY155227）. The coding sequence of scGH was cloned from pituitary of S. chuatsi by using RT-PCR and the construction of its eukaryotic expression vector was studied. The result showed that the entire open reading flame of the scGH was 615 bp and codified 204 amino acid residues. The sequence identity between the S. chuatsi GH and the S. kneri GH cDNA order in the GenBank was 99. 84%. The scGH cDNA order was inserted into Eukaryotic expression vec- tor（pYES2/CT） and was proved after selecting, digesting and sequencing.